Cyclic nucleotide phosphodiesterase of Dictyostelium discoideum and its glycoprotein inhibitor: structure and expression of their genes.
Cyclic nucleotide phosphodiesterase of Dictyostelium discoideum and its glycoprotein inhibitor: structure and expression of their genes.
复制标题
盘基网柄菌环核苷酸磷酸二酯酶及其糖蛋白抑制剂：其基因的结构和表达。
DOI：
10.1002/dvg.1020120118
复制
发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Kessin,RH
中科院分区：
文献类型：
--
作者：
Franke,J;Faure,M;Wu,L;Hall,AL;Podgorski,GJ;Kessin,RH
编码环核苷酸磷酸二酯酶（PD）和PD抑制糖蛋白（PDI）的基因已被克隆和表征。PDI基因是一个1.6 kb的基因组片段，包括含有两个小内含子的编码序列和510个核苷酸的5′端非翻译序列。从推导的氨基酸序列，我们预测的蛋白质的分子量（MW）为26,000，与以前的数据一致，含有15%的半胱氨酸残基。基因组Southern杂交分析表明，只有一个基因编码的抑制剂。北方印迹分析显示0.95 kb的单一转录物。PDI基因在发育早期表达，在聚集后几乎没有剩余的转录物。cAMP的存在阻止了PDI mRNA的出现，但当cAMP被去除时，转录物在30分钟内出现。当cAMP应用于表达PDI的细胞时，转录物消失，半衰期小于30分钟。D. discoideum转录成三种mRNA：生长特异性的1.9 kb mRNA、聚集特异性的2.4 kb mRNA和晚期发育特异性的2.2 kb mRNA。2.2 kb的mRNA也对前茎细胞具有特异性，并由分化诱导因子诱导。所有三种mRNA都含有相同的编码序列，仅在5′端非编码序列上有所不同。每个mRNA是从一个不同的启动子转录，并通过使用氯霉素酰基转移酶基因作为一个报告，我们已经表明，每个启动子显示其同源mRNA相同的调节。用PD基因转化野生型菌株会导致PD过表达，这会加速聚集并阻断随后的细胞分化和模式形成。
The genes coding for the cyclic nucleotide phosphodiesterase (PD) and the PD inhibitory glycoprotein (PDI) have been cloned and characterized. The PDI gene was isolated as a 1.6 kb genomic fragment, which included the coding sequence containing two small introns and 510 nucleotides of non‐translated 5′ sequence. From the deduced amino acid sequence we predict a protein with a molecular weight (MW) of 26,000 that, in agreement with previous data, contains 15% cysteine residues. Genomic Southern blot analysis indicates that only one gene encodes the inhibitor. Northern blot analysis shows a single transcript of 0.95 kb. The PDI gene is expressed early in development with little transcript remaining following aggregation. The appearance of PDI mRNA is prevented by the presence of cAMP, but when cAMP is removed the transcript appears within 30 minutes. When cAMP is applied to cells expressing PDI the transcript disappears with a half‐life of less than 30 minutes. The PD gene ofD. discoideumis transcribed into three mRNAs: a 1.9 kb mRNA specific for growth, a 2.4 kb mRNA specific for aggregation, and a 2.2 kb mRNA specific for late development. The 2.2 kb mRNA is also specific for prestalk cells, and is induced by differentiation‐inducing factor. All three mRNAs contain the same coding sequence, and differ only in their 5′ non‐coding sequences. Each mRNA is transcribed from a different promoter, and by using the chloramphenicol acyltransferase gene as a reporter, we have shown that each promoter displays the same regulation as its cognate mRNA. Transformation of wild‐type strains with the PD gene causes PD overexpression which accelerates aggregation and blocks subsequent cell differentiation and pattern formation.